LOCUS       sequence-208717-        8095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses pentamerised full-length protein ectodomain with no 
            N-linked glycans in mammalian cells. Contains a C-terminal rat Cd4 
            d3+4 tag, COMP pentamerisation domain, flag- and 6xHis tags. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8095)
  AUTHORS   Knockel J, Dundas K, Yang ASP, Galaway F, Metcalf T, Gemert GV, 
            Sauerwein RW, Rayner JC, Billker O, Wright GJ
  TITLE     Systematic Identification of Plasmodium Falciparum Sporozoite 
            Membrane Protein Interactions Reveals an Essential Role for the p24 
            Complex in Host Infection.
  JOURNAL   Mol Cell Proteomics. 2021;20:100038. doi: 10.1074/mcp.RA120.002432. 
            Epub 2021 Jan 27.
  PUBMED    33515807
REFERENCE   2  (bases 1 to 8095)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell 
            Proteomics."; date: "2021"; volume: "20:100038. doi"; pages: " 
            10.1074/mcp.RA120.002432. Epub 2021 Jan 27"
COMMENT     Sequence Label: PF3D7_0713700-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8095
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     310..329
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      352..361
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1627..2412
                     /codon_start=1
                     /gene="bla(M)"
                     /product="beta-lactamase lacking the signal sequence"
                     /label=bla(M)
                     /note="allows cytosolic expression of beta-lactamase"
                     /translation="PETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMST
                     FKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMS
                     DNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATT
                     LRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAA
                     LGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     CDS             2419..2484
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2491..2508
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2546..2601
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2600..2619)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4837..4855)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4923..5027
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5028..5096,5097..5888)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6059..6647
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6548..6567
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6801..6818
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6935..6956
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6971..6976,6977..6994,6995..7001)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7009..7025
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     7014..7036
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7033..7049
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7033..7049
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7320..7699
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7700..7903
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7854..7874
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7900..7924
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 ctcgagaaag gcgtctaacc agtcacagtc gcaaggtagg ctgagcaccg tggcgggcgg
       61 cagcgggtgg cggtcggggt tgtttctggc ggaggtgctg ctgatgatgt aattaaagta
      121 ggcggtcttg agacggcgga tggtcgaggt gaggtgtggc aggcttgaga tccagctgtt
      181 ggggtgagta ctccctctca aaagcgggca ttacttctgc gctaagattg tcagtttcca
      241 aaaacgagga ggatttgata ttcacctggc ccgatctggc catacacttg agtgacaatg
      301 acatccactt tgcctttctc tccacaggtg tccactccca ggtccaagtt tgccaccatg
      361 gagtttcaga cccaggtact catgtccctg ctgctctgca tgtctggtgc ggccgccgac
      421 atcatcatcg tggacagcaa caaggacatc cagggcttcg accccaacag cgtgctgaac
      481 ctggccgaga acgtgttcaa catcttcctg gacaccaaga ccaacaacgc cgatatcaag
      541 accgtgtaca gcagcatctt cagcaacagc aaagtgacca tcgacggcac ctgtagcagc
      601 atgaaggaac tgatcaaggg cagaagcctg aagtacctgg acgagctggg caccgaggaa
      661 tacgtgaaca gcgacgatgt gttcaacttc gccatcttcg tgtacaccgt gaacggccag
      721 cccgatatca tccagcaggt ctacaagaag caccccaact acatcagcta catcatcacc
      781 tgtaaagaga gcgagctgtt cgacaacatc gaggacgcca aggacaacaa gaatgtgttc
      841 cacaccaacc ccggcgcgcc gtcgacctcc atcacggcct ataagagtga gggggagtca
      901 gcggagttct ccttcccact caaccttgga gaggaaagcc tgcagggaga gttgagatgg
      961 aaggcagaga aggctccttc ttcccagtcc tggatcacct tctccctaaa gaaccaaaag
     1021 gtgtctgtgc agaagtctac tagcaacccc aagttccagc tgtccgaaac gctcccactc
     1081 acccttcaga taccccaggt ctcccttcag tttgctggtt ctggcaacct gaccctgact
     1141 ctggacagag ggatactgta tcaggaagtg aacctggtgg tgatgaaagt gactcagccc
     1201 gacagcaaca ctttgacctg tgaggtgatg ggacccacct cacccaagat gagactgatc
     1261 ttgaagcagg agaatcagga ggccagggtc tccaggcagg agaaagtgat tcaagtgcag
     1321 gcccctgaag caggggtgtg gcaatgtcta ctgagtgaag gtgaagaggt caagatggac
     1381 tccaagatcc aggttttatc caaagggttg aattccggcg gcggctccgg aggcggtacc
     1441 ggtggcggcg acctagcccc acagatgctt cgagaactcc aggagactaa tgcggcgctg
     1501 caagacgtga gagagctctt gcgacagcag gtcaaggaga tcaccttcct gaagaatacg
     1561 gtgatggaat gtgacgcctg cggtctagac agaaacctgc ctccactagc acctctgggc
     1621 ccacacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1681 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1741 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     1801 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1861 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1921 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1981 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     2041 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     2101 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     2161 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2221 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2281 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2341 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2401 attaagcatt ggggccctga ctacaaagac catgacggtg attataaaga tcatgacatc
     2461 gattacaagg atgacgatga caagggccct caccaccacc atcaccacta attaattaag
     2521 gatcccccga cctcgacctc tggctaataa aggaaattta ttttcattgc aatagtgtgt
     2581 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttggtcgag
     2641 atccctcgga gatctctagc tagaggatcg atccccgccc cggacgaact aaacctgact
     2701 acgacatctc tgccccttct tcgcggggca gtgcatgtaa tcccttcagt tggttggtac
     2761 aacttgccaa ctgggccctg ttccacatgt gacacggggg gggaccaaac acaaaggggt
     2821 tctctgactg tagttgacat ccttataaat ggatgtgcac atttgccaac actgagtggc
     2881 tttcatcctg gagcagactt tgcagtctgt ggactgcaac acaacattgc ctttatgtgt
     2941 aactcttggc tgaagctctt acaccaatgc tgggggacat gtacctccca ggggcccagg
     3001 aagactacgg gaggctacac caacgtcaat cagaggggcc tgtgtagcta ccgataagcg
     3061 gaccctcaag agggcattag caatagtgtt tataaggccc ccttgttaac cctaaacggg
     3121 tagcatatgc ttcccgggta gtagtatata ctatccagac taaccctaat tcaatagcat
     3181 atgttaccca acgggaagca tatgctatcg aattagggtt agtaaaaggg tcctaaggaa
     3241 cagcgatatc tcccacccca tgagctgtca cggttttatt tacatggggt caggattcca
     3301 cgagggtagt gaaccatttt agtcacaagg gcagtggctg aagatcaagg agcgggcagt
     3361 gaactctcct gaatcttcgc ctgcttcttc attctccttc gtttagctaa tagaataact
     3421 gctgagttgt gaacagtaag gtgtatgtga ggtgctcgaa aacaaggttt caggtgacgc
     3481 ccccagaata aaatttggac ggggggttca gtggtggcat tgtgctatga caccaatata
     3541 accctcacaa accccttggg caataaatac tagtgtagga atgaaacatt ctgaatatct
     3601 ttaacaatag aaatccatgg ggtggggaca agccgtaaag actggatgtc catctcacac
     3661 gaatttatgg ctatgggcaa cacataatcc tagtgcaata tgatactggg gttattaaga
     3721 tgtgtcccag gcagggacca agacaggtga accatgttgt tacactctat ttgtaacaag
     3781 gggaaagaga gtggacgccg acagcagcgg actccactgg ttgtctctaa cacccccgaa
     3841 aattaaacgg ggctccacgc caatggggcc cataaacaaa gacaagtggc cactcttttt
     3901 tttgaaattg tggagtgggg gcacgcgtca gcccccacac gccgccctgc ggttttggac
     3961 tgtaaaataa gggtgtaata acttggctga ttgtaacccc gctaaccact gcggtcaaac
     4021 cacttgccca caaaaccact aatggcaccc cggggaatac ctgcataagt aggtgggcgg
     4081 gccaagatag gggcgcgatt gctgcgatct ggaggacaaa ttacacacac ttgcgcctga
     4141 gcgccaagca cagggttgtt ggtcctcata ttcacgaggt cgctgagagc acggtgggct
     4201 aatgttgcca tgggtagcat atactaccca aatatctgga tagcatatgc tatcctaatc
     4261 tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     4321 tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc tatcctaatc
     4381 tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatc
     4441 tgtatccggg tagcatatgc tatcctaata gagattaggg tagtatatgc tatcctaatt
     4501 tatatctggg tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata
     4561 tctgggtagc atatgctatc ctaatctata tctgggtagc ataggctatc ctaatctata
     4621 tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc ctaatttata
     4681 tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata
     4741 tctgggtagt atatgctatc ctaatctgta tccgggtagc atatgctatc ctcatgataa
     4801 gctgtcaaac atgagaattt tcttgaagac gaaagggcct cgtgatacgc ctatttttat
     4861 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     4921 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4981 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     5041 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5101 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5161 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5221 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     5281 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5341 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5401 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5461 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5521 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
     5581 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5641 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5701 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5761 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5821 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5881 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5941 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6001 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6061 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6121 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6181 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6241 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6301 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6361 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6421 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6481 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6541 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6601 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6661 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6721 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     6781 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     6841 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     6901 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     6961 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     7021 aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag ctagaggtcg
     7081 accaattctc atgtttgaca gcttatcatc gcagatccgg gcaacgttgt tgccattgct
     7141 gcaggcgcag aactggtagg tatggaagat ctatacattg aatcaatatt ggcaattagc
     7201 catattagtc attggttata tagcataaat caatattggc tattggccat tgcatacgtt
     7261 gtatctatat cataatatgt acatttatat tggctcatgt ccaatatgac cgccatgttg
     7321 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     7381 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     7441 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     7501 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     7561 agtgtatcat atgccaagtc cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     7621 gcattatgcc cagtacatga ccttacggga ctttcctact tggcagtaca tctacgtatt
     7681 agtcatcgct attaccatgg tgatgcggtt ttggcagtac accaatgggc gtggatagcg
     7741 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     7801 gcaccaaaat caacgggact ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat
     7861 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca
     7921 gatcctcact ctcttccgca tcgctgtctg cgagggccag ctgttgggct cgcggttgag
     7981 gacaaactct tcgcggtctt tccagtactc ttggatcgga aacccgtcgg cctccgaacg
     8041 gtactccgcc accgagggac ctgagcgagt ccgcatcgac cggatcggaa aacct
//
